Ultrasound-Targeted Microbubble Destruction Delivery of Insulin-Like Growth Factor 1 cDNA and Transforming Growth Factor Beta Short Hairpin RNA Enhances Tendon Regeneration and Inhibits Scar Formation In Vivo

Abstract

Ultrasound-targeted microbubble destruction (UTMD), which has been successfully used for the treatment of many diseases, offers a promising noninvasive approach for target-specific gene delivery. This study investigated the UTMD delivery of insulin-like growth factor 1 (IGF-1) cDNA and transforming growth factor beta (TGF-β) short hairpin RNA for Achilles tendon injury in rats. Briefly, 168 rats with an injured Achilles tendon were randomly divided into seven groups: (1) IGF-1 + UTMD, (2) TGF-β + UTMD, (3) IGF-1 + TGF-β + UTMD, (4) control, (5) IGF-1, (6) TGF-β, and (7) IGF-1 + TGF-β. At 2, 4, 8, and 12 weeks post treatment, six rats from each group were euthanized. IGF-1 expression and TGF-β expression were evaluated using an adhesion index score, pathological examination, quantitative real-time reverse transcription polymerase chain reaction, Western blotting, and biomechanical measurement. The lowest adhesion index score, the lightest inflammation, the highest 4,6-diamidino-2-phenylindole nuclear counter signals, the highest IGF-1 expression, and the lowest TGF-β expression were observed in group 3 (p < 0.05). Furthermore, higher expression of IGF-1 mRNA was observed in groups 1 and 3, while lower expression of TGF-β mRNA was observed in groups 2 and 3 (p < 0.05). The UTMD groups showed a higher transfection efficiency than the groups without UTMD. Downregulation of type III collagen and upregulation of type I collagen were observed in groups 1-3. Moreover, during weeks 4, 8, and 12, greater maximum load and tensile stress were observed in group 3 compared to the other groups (p < 0.05), while the highest tendon stiffness was observed in week 12 (p < 0.05). To conclude, the results suggest that UTMD delivery of IGF-1 and TGF-β offers a promising treatment approach for tendon injury in vivo.